Agilent Protocol
DNA
All reagents must be at room temperature
To get started, add 25ul Dye into the entire matrix tube, filter (tube provided)
(Note: this solution is good for up to one month)
Chip holds 12 samples
Note: RNA work, syringe clip at top; DNA work, syringe clip at bottom
1. Prime the Chip with Matrix/dye solution through the wells (inspect to see no bubbles)
a. 1st G1 well: 9ul Matrix/dye solution (touch tip, only soft stop, no air)
b. Snap down, inject
c. Wait 1 minute
d. Release syringe. Let it come up on its own (a few seconds)
2. Other 2 G wells: 9ul Matrix/dye solution
3. Ladder well: 5ul Matrix/dye solution
4. Sample wells: 5ul marker into each
5. Ladder well: 1ul ladder
6. Sample wells: 1ul sample into individual wells
7. Shake at 2400 for 1 minute
8. Computer
a. Open Agilent
b. Assay; dsDNA; DNA 500
c. Start
Assay takes about 20 minutes
RNA
Note: NO HARD STOPS!
1. Prepare matrix (red cap)- column purify 550ul- should last 1 month- spin at 3000gx10min. If
doing only a few samples you can column purify about 200ul
2. Prepare DyeMatrix- to 65ul filtered matrix add 1ul dye (blue cap)- vortex. Spin 10 min at 13000g.
Keep away from bottom when pipetting.
3. The ladder is purchased separately (in -80 freezer). Thaw the ladder and heat 1.5ul in a new
tube to 65-70 degrees (heat block) for 2 minutes. Let sit on ice until needed.
4. Put syringe clip at the top
5. Add 9ul of DyeMatrix into G1. Inject- wait 30 seconds. Release and let syringe come up.
6. Add 9ul of DyeMatrix into the other two G wells.
7. Ladder well: Add 5ul marker (green cap)
8. Sample wells: add 5ul marker
9. Ladder well: Add 1ul ladder (first heat at 70 degrees for 2 minutes, place on ice)
10. Sample well: add 1ul of each sample to well
11. Shake 2400 for 1 minute
12. Computer
a. Assay>RNA
b. Eukaryote total RNA nano
c. Start
13. Clean after with 1 minute RNA zap, 1 minute RNAse free water